We examined the role of arbuscular mycorrhizal fungi (AMF) in ecosystems using soil aggregate stability and C and N storage as representative ecosystem processes. We utilized a wide gradient in AMF abundance, obtained through long-term (17 and 6 years) large-scale field manipulations. Burning and N-fertilization increased soil AMF hyphae, glomalin-related soil protein (GRSP) pools and water-stable macroaggregates while fungicide applications reduced AMF hyphae, GRSP and water-stable macroaggregates. We found that AMF abundance was a surprisingly dominant factor explaining the vast majority of variability in soil aggregation. This experimental field study, involving long-term diverse management practices of native multispecies prairie communities, invariably showed a close positive correlation between AMF hyphal abundance and soil aggregation, and C and N sequestration. This highly significant linear correlation suggests there are serious consequences to the loss of AMF from ecosystems.